1887

Abstract

and ureases are nickel-requiring metallo-enzymes that hydrolyse urea to NH and CO. In both and in an model of urease activity, a high affinity nickel transporter, NixA, is required for optimal urease activity, whereas the urea-dependent UreR positive transcriptional activator governs optimal urease expression in . The gene is a flagellar biosynthesis and regulatory gene that modulates urease activity in the model of urease activity. All mutants of eight strains of were non-motile and five had a strain-dependent alteration in urease activity. The gene decreased urease activity 15-fold when expressed in containing the urease locus and the gene; this was reversed by disruption of . The gene decreased transcription. also decreased urease activity three-fold in containing the urease locus in a urea- and UreR-dependent fashion. Here the gene repressed the urease promoter. Thus, FlbA decreased urease activity of both and , but through distinct mechanisms. wild-type strain SS1 colonised gerbils at a mean of 5.4×105mucfu/g of antrum and caused chronic gastritis and lesions in the antrum. In contrast, the mutant did not colonise five of six gerbils and caused no lesions, indicating that motility mediated by was required for colonisation. Because FlbA regulates flagellar biosynthesis and secretion, as well as forming a structural component of the flagellar secretion apparatus, two seemingly unrelated virulence attributes, motility and urease, may be coupled in and and possibly also in other motile, ureolytic bacteria.

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2002-11-01
2019-11-22
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References

  1. Blaser MJ. Helicobacter pylori and the pathogenesis of gastroduodenal inflammation. J Infect Dis 1990;; 161: 626–633.[CrossRef]
    [Google Scholar]
  2. Buck GE, Gourley WK, Lee WK, Subramanyan JM, Latimer JM, Di Nuzzo AR. Relation of Campylobacter pyloridis to gastritis and peptic ulcers. J Infect Dis 1986;; 153: 664–669.[CrossRef]
    [Google Scholar]
  3. International Agency for Cancer Research. IARC Monographs on the evaluations of cancer risks to humans. Lyon, World Health Organization. 1994:; 177–240.
  4. Parsonnet J, Friedman GD, Vandersteen DP.et al. Helicobacter pylori infection and the risk of gastric carcinoma. N Engl J Med 1991;; 325: 1127–1131.[CrossRef]
    [Google Scholar]
  5. McGee DJ, Mobley HLT. Mechanisms of Helicobacter pylori infection: bacterial factors. In: Westblom TU, Czinn SJ, Nedrud JG (eds) Gastroduodenal disease and Helicobacter pylori: pathophysiology, diagnosis and treatment. Current topics in microbiology and immunology, vol 24.Berlin, Springer-Verlag. 1999:; 155–180.
  6. McGee DJ, Mobley HLT. Pathogenesis of Helicobacter pylori infection. Cur Opin Gastroenterol 2000;; 16: 24–31.[CrossRef]
    [Google Scholar]
  7. Mobley HLT, Island MD, Hausinger RP. Molecular biology of microbial ureases. Microbiol Rev 1995;; 59: 451–480.
    [Google Scholar]
  8. Bauerfeind P, Garner RM, Mobley HLT. Allelic exchange mutagenesis of nixA in Helicobacter pylori results in reduced nickel transport and urease activity. Infect Immun 1996;; 64: 2877–2880.
    [Google Scholar]
  9. McGee DJ, May CA, Garner RM, Himpsl JM, Mobley HLT. Isolation of Helicobacter pylori genes that modulate urease activity. J Bacteriol 1999;; 181: 2477–2484.
    [Google Scholar]
  10. Mobley HLT, Garner RM, Bauerfeind P. Helicobacter pylori nickel-transport gene nixA: synthesis of catalytically active urease in Escherichia coli independent of growth conditions. Mol Microbiol 1995;; 16: 97–109.[CrossRef]
    [Google Scholar]
  11. Ferrero RL, Hazell SL, Lee A. The urease enzymes of Campylobacter pylori and a related bacterium. J Med Microbiol 1988;; 27: 33–40.[CrossRef]
    [Google Scholar]
  12. Hu L-T, Mobley HLT. Expression of catalytically active recombinant Helicobacter pylori urease at wild-type levels in Escherichia coli. Infect Immun 1993;; 61: 2563–2569.
    [Google Scholar]
  13. Slonczewski JL, McGee DJ, Phillips J, Kirkpatrick C, Mobley HLT. pH-dependent protein profiles of Helicobacter pylori analyzed by two-dimensional gels. Helicobacter 2000;; 5: 240–247.[CrossRef]
    [Google Scholar]
  14. Scott DR, Weeks D, Hong C, Posius S, Melchers K, Sachs G. The role of internal urease in acid resistance of Helicobacter pylori. Gastroenterology 1998;; 114: 58–70.[CrossRef]
    [Google Scholar]
  15. Weeks DL, Eskandari S, Scott DR, Sachs G. A H+-gated urea channel: the link between Helicobacter pylori urease and gastric colonization. Science 2000;; 287: 482–485.[CrossRef]
    [Google Scholar]
  16. van Vliet AHM, Kuipers EJ, Waidner B.et al. Nickel-responsive induction of urease expression in Helicobacter pylori is mediated at the transcriptional level. Infect Immun 2001;; 69: 4891–4897.[CrossRef]
    [Google Scholar]
  17. Mobley HLT, Warren JW. Urease-positive bacteriuria and obstruction of long-term urinary catheters. J Clin Microbiol 1987;; 25: 2216–2217.
    [Google Scholar]
  18. Róz2alski A, Sidorczyk Z, Kotełko K. Potential virulence factors of Proteus bacilli. Microbiol Mol Biol Rev 1997;; 61: 65–89.
    [Google Scholar]
  19. Warren JW, Tenney JH, Hoopes JM, Muncie HL, Anthony WC. A prospective microbiologic study of bacteriuria in patients with chronic indwelling urethral catheters. J Infect Dis 1982;; 146: 719–723.[CrossRef]
    [Google Scholar]
  20. Jones BD, Lockatell CV, Johnson DE, Warren JW, Mobley HLT. Construction of a urease-negative mutant of Proteus mirabilis: analysis of virulence in a mouse model of ascending urinary tract infection. Infect Immun 1990;; 58: 1120–1123.
    [Google Scholar]
  21. Nicholson EB, Concaugh EA, Foxall PA, Island MD, Mobley HLT. Proteus mirabilis urease: transcriptional regulation by UreR. J Bacteriol 1993;; 175: 465–473.
    [Google Scholar]
  22. Jones BD, Mobley HLT. Proteus mirabilis urease: genetic organization, regulation, and expression of structural genes. J Bacteriol 1988;; 170: 3342–3349.
    [Google Scholar]
  23. Alm RA, Ling L-SL, Moir DT.et al. Genomic-sequence comparison of two unrelated isolates of the human gastric pathogen Helicobacter pylori. Nature 1999;; 397: 176–180.[CrossRef]
    [Google Scholar]
  24. Tomb J-F, White O, Kerlavage AR.et al. The complete genome sequence of the gastric pathogen Helicobacter pylori. Nature 1997;; 388: 539–547.[CrossRef]
    [Google Scholar]
  25. Mobley HLT, Cortesia MJ, Rosenthal LE, Jones BD. Characterization of urease from Campylobacter pylori. J Clin Microbiol 1988;; 26: 831–836.
    [Google Scholar]
  26. Island MD, Mobley HLT. Proteus mirabilis urease: operon fusion and linker insertion analysis of ure gene organization, regulation, and function. J Bacteriol 1995;; 177: 5653–5660.
    [Google Scholar]
  27. Schmitz A, Josenhans C, Suerbaum S. Cloning and characterization of the Helicobacter pylori flbA gene, which encodes for a membrane protein involved in coordinated expression of flagellar genes. J Bacteriol 1997;; 179: 987–997.
    [Google Scholar]
  28. Plano GV, Barve SS, Straley SC. LcrD, a membrane-bound regulator of the Yersinia pestis low-calcium response. J Bacteriol 1991;; 173: 7293–7303.
    [Google Scholar]
  29. Andrews GP, Maurelli AT. mxiA of Shigella flexneri 2a, which facilitates export of invasion plasmid antigens, encodes a homolog of the low-calcium response protein, LcrD, of Yersinia pestis. Infect Immun 1992;; 60: 3287–3295.
    [Google Scholar]
  30. Galán JE, Ginocchio C, Costeas P. Molecular and functional characterization of the Salmonella invasion gene invA: homology of InvA to members of a new protein family. J Bacteriol 1992;; 174: 4338–4349.
    [Google Scholar]
  31. Ginocchio CC, Galán JE. Functional conservation among members of the Salmonella typhimurium InvA family of proteins. Infect Immun 1995;; 63: 729–732.
    [Google Scholar]
  32. Plano GV, Straley SC. Multiple effects of lcrD mutations in Yersinia pestis. J Bacteriol 1993;; 175: 3536–3545.
    [Google Scholar]
  33. Fleiszig SMJ, Arora SK, Van R, Ramphal R. FlhA, a component of the flagellum assembly apparatus of Pseudomonas aeruginosa, plays a role in internalization by corneal epithelial cells. Infect Immun 2001;; 69: 4931–4937.[CrossRef]
    [Google Scholar]
  34. Eaton KA, Morgan DR, Krakowka S. Motility as a factor in the colonisation of gnotobiotic piglets by Helicobacter pylori. J Med Microbiol 1992;; 37: 123–127.[CrossRef]
    [Google Scholar]
  35. Eaton KA, Suerbaum S, Josenhans C, Krakowka S. Colonization of gnotobiotic piglets by Helicobacter pylori deficient in two flagellin genes. Infect Immun 1996;; 64: 2445–2448.
    [Google Scholar]
  36. Kim JS, Chang JH, Chung SI, Yum JS. Molecular cloning and characterization of the Helicobacter pylori fliD gene, an essential factor in flagellar structure and motility. J Bacteriol 1999;; 181: 6969–6976.
    [Google Scholar]
  37. Iwao E, Hirayama F, Takagai S, Yokoyama Y, Ikeda Y. Virulence factors of Helicobacter pylori affecting its gastric colonization in Mongolian gerbils. J Gastroenterol 1999;; 34 Suppl XI: 47–54.
    [Google Scholar]
  38. Testerman TL, McGee DJ, Mobley HLT. Helicobacter pylori growth and urease detection in the chemically-defined medium Ham's F-12 nutrient mixture. J Clin Microbiol 2001;; 39: 3842–3850.[CrossRef]
    [Google Scholar]
  39. Ausubel FM, Brent R, Kingston RE et al. (eds). Current protocols in molecular biology. New York, John Wiley & Sons. 1995.
  40. Sambrook, J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor, NY, Cold Spring Harbor Laboratory. 1989.
  41. McAllister CF, Stephens DS. Analysis in Neisseria meningitidis and other Neisseria species of genes homologous to the FKBP immunophilin family. Mol Microbiol 1993;; 10: 13–23.[CrossRef]
    [Google Scholar]
  42. McGee DJ, Radcliff FJ, Mendz GL, Ferrero RL, Mobley HLT. The Helicobacter pylori rocF is required for arginase activity and acid protection in vitro but is not essential for in vivo colonization of mice or for urease activity. J Bacteriol 1999;; 181: 7314–7322.
    [Google Scholar]
  43. Miller JH. Experiments in molecular genetics. Cold Spring Harbor, NY, Cold Spring Harbor Laboratory. 1977:; 352–354.
  44. Josenhans C, Eaton KA, Thevenot T, Suerbaum S. Switching of flagellar motility in Helicobacter pylori by reversible length variation of a short homopolymeric sequence repeat in fliP, a gene encoding a basal body protein. Infect Immun 2000;; 68: 4598–4603.[CrossRef]
    [Google Scholar]
  45. Hu L-T, Foxall PA, Russell R, Mobley HLT. Purification of recombinant Helicobacter pylori urease apoenzyme encoded by ureA and ureB. Infect Immun 1992;; 60: 2657–2666.
    [Google Scholar]
  46. Nixon DE, Moyer TP, Squillace DP, McCarthy JT. Determination of serum nickel by graphite furnace atomic absorption spectrometry with Zeeman-effect background correction: values in a normal population and a population undergoing dialysis. Analyst 1989;; 114: 1671–1674.[CrossRef]
    [Google Scholar]
  47. Sunderman FW, Crisostomo MC, Reid MC, Hopfer SM, Nomoto S. Rapid analysis of nickel in serum and whole blood by electrothermal atomic absorption spectrophotometry. Ann Clin Lab Sci 1984;; 14: 232–241.
    [Google Scholar]
  48. Templeton DM, Sunderman FW, Herber RF. Tentative reference values for nickel concentrations in human serum, plasma, blood, and urine: evaluation according to the TRACY protocol. Sci Total Environ 1994;; 148: 243–251.[CrossRef]
    [Google Scholar]
  49. Sunderman FW, Hopfer SM, Crisostomo MC, Stoeppler M. Rapid analysis of nickel in urine by electrothermal atomic absorption spectrophotometry. Ann Clin Lab Sci 1986;; 16: 219–230.
    [Google Scholar]
  50. Soriano A, Hausinger RP. GTP-dependent activation of urease apoprotein in complex with the UreD, UreF, and UreG accessory proteins. Proc Natl Acad Sci USA 1999;; 96: 11140–11144.[CrossRef]
    [Google Scholar]
  51. Akopyants NS, Fradkov A, Diatchenko L.et al. PCR-based subtractive hybridization and differences in gene content among strains of Helicobacter pylori. Proc Natl Acad Sci USA 1998;; 95: 13108–13113.[CrossRef]
    [Google Scholar]
  52. Achtman M, Azuma T, Berg DE.et al. Recombination and clonal groupings within Helicobacter pylori from different geographical regions. Mol Microbiol 1999;; 32: 459–470.[CrossRef]
    [Google Scholar]
  53. Kersulyte D, Chalkauskas H, Berg DE. Emergence of recombinant strains of Helicobacter pylori during human infection. Mol Microbiol 1999;; 31: 31–43.[CrossRef]
    [Google Scholar]
  54. Suerbaum S, Maynard Smith J, Bapumia K.et al. Free recombination within Helicobacter pylori. Proc Natl Acad Sci USA 1998;; 95: 12619–12624.[CrossRef]
    [Google Scholar]
  55. Eaton KA, Kersulyte D, Mefford M, Danon SJ, Krakowka S, Berg DE. Role of Helicobacter pylori cag region genes in colonization and gastritis in two animal models. Infect Immun 2001;; 69: 2902–2908.[CrossRef]
    [Google Scholar]
  56. Hendricks JK, Mobley HLT. Helicobacter pylori ABC transporter: effect of allelic exchange mutagenesis on urease activity. J Bacteriol 1997;; 179: 5892–5902.
    [Google Scholar]
  57. Dunn BE, Campbell GP, Perez-Perez GI, Blaser MJ. Purification and characterization of urease from Helicobacter pylori. J Biol Chem 1990;; 265: 9464–9469.
    [Google Scholar]
  58. Gygi D, Bailey MJ, Allison C, Hughes C. Requirement for FlhA in flagella assembly and swarm-cell differentiation by Proteus mirabilis. Mol Microbiol 1995;; 15: 761–769.
    [Google Scholar]
  59. Nakamura H, Yoshiyama H, Takeuchi H, Mizote T, Okita K, Nakazawa T. Urease plays an important role in the chemotactic motility of Helicobacter pylori in a viscous environment. Infect Immun 1998;; 66: 4832–4837.
    [Google Scholar]
  60. Yoshiyama H, Nakamura H, Kimoto M, Okita K, Nkazawa T. Chemotaxis and motility of Helicobacter pylori in a viscous environment. J Gastroenterol 1999;; 34 Suppl 11: 18–23.[CrossRef]
    [Google Scholar]
  61. Yoshiyama H, Nakazawa T. Unique mechanism of Helicobacter pylori for colonizing the gastric mucus. Microbes Infect 2000;; 2: 55–60.[CrossRef]
    [Google Scholar]
  62. Simons RW, Houman F, Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene 1987;; 53: 85–96.[CrossRef]
    [Google Scholar]
  63. Wang RF, Kushner SR. Construction of versatile low-copy-number vectors for cloning, sequencing and gene expression in Escherichia coli. Gene 1991;; 100: 195–199.[CrossRef]
    [Google Scholar]
  64. Gherardini FC, Hobbs MM, Stam M LV, Bassford PJ. Complementation of an Escherichia coli proC mutation by a gene cloned from Treponema pallidum. J Bacteriol 1990;; 172: 2996–3002.
    [Google Scholar]
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